These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Urine Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 250 µL to 1 mL.
Urine Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 2 mL to 10 mL.
Urine Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 11 mL to 30 mL.
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| Kit Specifications | |
| Minimum Urine Input | 2 mL |
| Maximum Urine Input | 10 mL |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50 – 100 µL |
| Time to Complete 10 Purifications | 35 – 40 minutes |
| Average Yields* | Variable depending on the specimen |
*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.
| Component | Cat. 59200 (50 preps) | Cat. 59300 (25 preps) | Cat. 59400 (15 preps) |
|---|---|---|---|
| Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
| ExoC Buffer | 8 mL | 30 mL | 50 mL |
| ExoR Buffer | 12 mL | 12 mL | 12 mL |
| Lysis Buffer A | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
| Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Columns | 50 | 25 | 15 |
| Mini Spin Columns | 100 | 50 | 30 |
| Collection Tubes | 100 | 50 | 30 |
| Elution Tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |
The BK virus is a member of the polyomavirus family. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Once an individual has been infected the virus disseminates to the kidneys and the urinary tract where it remains for the lifetime of the individual. Infections with BK virus in immunocompromised or immunosupressed patients are much more severe and may involve renal dysfunction. In fact, in kidney transplant patients the immunosupressive drugs required for the transplant may allow the virus to replicate within the graft, resulting in a disease called BK virus nephropathy (BKVN). It is thought that 1-10% of renal transplant patients progress to BK virus nephropathy (BKVN) and up to 80% of these patients are reported to have lost their grafts. The onset of nephritis can occur as early as several days post-transplant to as late as 5 years. The mode of transmission of the virus is not clear, however it has been suggested that BKV may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. This virus can be diagnosed by BKV blood & urine testing, in addition to carrying out a biopsy in the kidneys. PCR techniques are now widely used to identify the virus.
BKV TaqMan PCR Kit, 100 reactions
BKV TaqMan PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM36550 (100 preps) | Cat. TM36510 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| BKV Primer & Probe Mix | 280 μL | 280 μL |
| BKV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.
Specifications
| Features | Specifications |
| Recommended application | Small amounts of nucleic acid isolation, viral nucleic acid from cell free samples |
| Preservation conditions | Room temperature |
| Stability | Up to 4 years |
| Filter membrane | High quality glass fiber filter GF/F, 2 layers |
| Membrane aperture | 0.7μm |
| Maximum binding yield of plasmid | 30 μg |
| Maximum yield of alcohol mediated Binding | 100 μg |
| Single liquid carrying capacity of column | 900 μl |
| Minimum elution volume | 80 μl |
| Withstand centrifugal force | 5,000 x g |
| Centrifuge | Low speed centrifuge, Swing out Rotor, can placed a height of 6.5cm square, (height of HiPure DNA Plate & 1.6ml Collection Plate: height, 6.2cm) |
Adsorption Mechanism
Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silica gel membrane, while other pollutants, mainly proteins, are removed by membrane washing.
Ordering information
| CAT.No. | Product Name | Package |
| C13130 | HiPure DNA Plate (2 x GF/F)with 1.6ml Collection Plate | 10/Bag |
| Item No. | Product Name | Membrane type/number of layers | Collection tubes | Plasmid DNA binding capacity (Physical adsorption) | gDNA/RNA binding capacity (Alcohol-mediated adsorption) | Minimum Elution volume | Liquid volume capacity |
| C13010 | HiPure DNA Nano Column | 2 layers GF/F | 2ml without cap | 5μg | 20μg | 10μl | 700μl |
| C13011 | HiPure DNA Micro Column | 3 layers GF/F | 2ml without cap | 10μg | 50μg | 15μl | 700μl |
| C13100 | HiPure DNA Mini Column I | 2 layers GF/B | 2ml without cap | 15μg | 100μg | 30μl | 700μl |
| C13110 | HiPure DNA Mini Column II | 4 layers GF/B | 2ml without cap | 35μg | 200μg | 50μl | 800μl |
| C13111 | HiPure RNA Mini Column | 3 layers GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
| C13112 | HiPure Viral Mini Column | 3 layers GF/F | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
| C13113 | HiPure CFDNA Mini Column | 3 layers GF/F,1 layer GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
| C13120 | HiPure DNA Midi Column | 4 layers GF/B | 15ml Centrifuge tube | 125μg | 1mg | 500μl | 4ml |
| C13121 | HiPure DNA Midi Column III | 8 layers GF/B | 15ml Centrifuge tube | 250μg | 1mg | 500μl | 4ml |
| C13122 | HiPure DNA Maxi Column | 4 layers GF/B | 50ml Centrifuge tube | 500μg | 5mg | 1000μl | 20ml |
| C13123 | HiPure DNA Maxi Column III | 8 layers GF/B | 50ml Centrifuge tube | 1mg | 5mg | 1000μl | 20ml |
| C13124 | HiPure DNA Maxi Column C | 8 layers GF/B | 50ml high speed Centrifuge tube | 1mg | 5mg | 700μl | 12ml |
| C13130 | HiPure DNA Plate | 2 layers GF/F | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
| C13131 | HiPure gDNA Plate | 2 layers GF/B | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.